Determination of Anomalin and Deltoin in Seseli resinosum by LC Combined with Chemometric Methods

Liquid chromatography-chemometric methods [LC-Partial least squares (LC-PLS), LC-principle component regression (LC-PCR) and LC-artificial neural network (LC-ANN)] were developed for the determination of (-)-anomalin (ANO) and deltoin (DEL) in the root and aerial part of Seseli resinosum Freyn et Sint. (Umbelliferae). Firstly, chemometric conditions were optimized by testing different mobile phases at various proportions of solvents with various flow rates in different wavelengths by using a normal phase column to obtain the best separation and recovery results. As a result, a mobile phase consisting of n-hexane and ethyl acetate (75:25 v/v) at a constant flow rate of 0.8 mL min-¹ on the above column system at ambient temperature were found to be the optimal chromatographic condition for good separation and determination of ANO and DEL in samples. Multichromatograms for the concentration set containing ANO and DEL compounds in the concentration range of 50-400 ng mL-¹ were obtained by using a diode array detector (DAD) system at selected wavelength sets, 300 (A), 310 (B), 320 (C), 330 (D) and 340 (E). Three LC-chemometric approaches were applied to the multichromatographic data to construct chemometric calibrations. As an alternative method, traditional LC at single wavelength was used for the analysis of the related compounds in the plant extracts. All of the methods were validated by analyzing various synthetic ANO-DEL mixtures. After the above step, traditional and chemometric LC methods were applied to the real samples consisting of extracts from roots and aerial parts of S. resinosum. The results obtained by LC-chemometric approaches were compared to each other and with those obtained by traditional LC.